Chocolates, compounds and spreads: A review on the use of oleogels, hydrogels and hybrid gels to reduce saturated fat content.

This study is a bibliometric analysis and literature review on the use of oleogels (OGs), hydrogels (HGs) and hybrid gels (HYGs) in chocolate, compounds and spreads with the aim of reducing the saturated fat in these products. The articles were selected by analyzing titles, keywords and abstracts in the Web of Science (WoS), Scopus and Google Scholar databases. Supplementary documents were obtained from government sources, including patent registrations. The theoretical and practical aspects were critically analyzed, highlighting the main points of agreement and disagreement between the authors. The results revealed a lack of regulations and official guidelines that widely allow the use of OGs, HGs and HYGs in chocolate confectionery products. The type and characteristics of raw materials affect the properties of products. Replacing cocoa butter (CB) with OGs, HGs or HYGs also affects texture, melting point and behavior, and nutritional aspects. These substitutions can result in products with better sensory acceptance and health benefits, such as reducing saturated fat and promoting cardiovascular health. However, it is important to find the ideal combination and proportions of components to obtain the desired properties in the final products.

Influence of cocoa varieties on carbohydrate composition and enzymatic activity of cocoa pulp.

In Brazil, after the witch's broom disease incidence, diverse cocoa hybrids were developed, and variations were reported on their composition and characteristics. Based on this, the present study aimed to evaluate the pulp composition of several cocoa hybrids in order to better understand these variations. Results show that cocoa pulp is composed, on average, of 76 % sugar, and a wide variation (20 %) was observed in sugar content between hybrids. Regarding the sugar profile, a prevalence of reducing sugars was observed. Pod origin also plays an important role in pulp composition, with variations between hybrids from Espírito Santo and Bahia states. In relation to the degree of ripeness, ripe pods showed higher fructose and glucose content, while unripe pods presented mainly sucrose. Similar to sugars, the cello-oligossacharides profile was influenced by the degree of pod ripeness and origin and most ripe samples presented mainly cellobiose, cellotriose and cellotetrose. In addition, the prebiotic potential of cocoa pulp was highlighted by cello-oligossacharides digestion assay which exhibited low rates of degradation. Varying enzymatic activity was observed amongst different pulp hybrids, with polyphenol oxidase showing a higher variation when compared to invertase and polygalacturonase ranging. This study shows that the pod hybrid, origin and ripening degree may change the cocoa pulp composition. Therefore, it is very important to understand and evaluate these variations, in order to obtain better results in pulp utilization either in cocoa fermentation or as a coproduct.

Varietal Influence on the Formation of Bioactive Amines during the Processing of Fermented Cocoa with Different Pulp Contents.

During cocoa processing, there can be the formation of bioactive amines, which are compounds that play relevant roles not only in plant development but also in human health. Thus, we aimed to investigate the presence and levels of bioactive amines during the processing of two important varieties of cocoa (PS 1319 and Parazinho). The seeds were fermented using five different pulp proportions: 100% (E1), 80% (E2), 60% (E3), and 0% (total pulp removal) (E4). The beans were fermented and dried on a farm following traditional procedures. Soon after, they were roasted and processed into chocolates with 60% cocoa in the laboratory. Bioactive amine contents were determined by ion-pair reversed-phase HPLC and fluorometric detection in the samples before, during, and after fermentation, after drying and roasting (nibs), and in the liquor and chocolate. The only amines found before processing in PS 1319 and Parazinho, respectively, in dry weight basis (dwb), were putrescine (pulp, 13.77 and 12.31; seed, 5.88 and 4.58) and serotonin (seed, 2.70 and 2.54). Fermentation was shorter for Parazinho (156 h) compared to PS 1319 (180 h). The changes in amines were affected by the cocoa variety. During drying, the presence of cadaverine stood out, appearing in all treatments of the PS 1319 variety, reaching 17.96 mg/kg dwb, and in two treatments of the Parazinho variety (100 and 60% pulp). During roasting, most of the amines decreased, except for phenylethylamine, which increased up to 2.47 mg/kg dwb for Parazinho and 1.73 mg/kg dwb for PS 1319. Most of the amines formed and built up (e.g., tyramine, putrescine, and cadaverine) during fermentation were not available or were at low levels in the nibs. Most of the amines found during processing did not reach the final product (chocolate), except for cadaverine in PS 1319 without pulp (7.54 mg/kg dwb). Finally, we confirmed how pulp content, processing, and variety influence the content of bioactive amines in cocoa and chocolate. These changes can be better demonstrated through a heatmap and principal component analysis.

Cupuassu from bean to bar: Sensory and hedonic characterization of a chocolate-like product.

The processing of cupuassu (Theobroma grandiflorum Schum) beans after fermentation gives a chocolate-like product, the cupulate. The high amount of pulp adhered to the seeds hinders the fermentation. Consequently, it is necessary to depulp the seeds to perform the process, even though the pulp contains important substrates for the formation of flavor precursors. To verify whether the complete or partial removal of the pulp influences the sensory characteristics of the product, fermentation was performed with three pulp concentrations (0, 7.5, and 15%) and two schemes of turning for aeration of the mass: fixed (R1) and according to the temperature (R2), in a total of six experiments (0R1, 0R2, 7.5R1, 7.5R2, 15R1 and 15R2). The beans were processed to obtain cupulates, which were submitted to tests performed with consumers, to express their preference and attributes (acceptance, purchase intent, Check All That Apply - CATA), and then to tests with a trained panel, the Quantitative Descriptive Profile (QDP) to characterize the samples. Both tests showed the consumers' perceptions that the cupulates have peculiar sensory characteristics. In the Consumer Test, through the Preference Mapping, all the samples of cupulates obtained from R1 conditions were preferred. In the penalty analysis, these same samples showed positive attributes that mask the negative attributes. Both CATA and QDP results showed that cupulate samples produced from seeds with a higher amount of pulp (15R1 and 15R2) had a higher number of positive mentions, for their fruity and floral flavors. The research also demonstrated that all samples gave the perception of an earthy taste, an important reason for consumer rejection, as well as a bad residual flavor. Thus, the results showed that the presence of the pulp in the fermentation environmental is important to the formation of flavor compounds and improving the sensory acceptance of the products.

Beetroot and radish powders as natural nitrite source for fermented dry sausages.

The aim of this study was to investigate the use of radish and beetroot powders as potential substitutes of nitrite in fermented dry sausages due to their high nitrate content (around 16,000 and 14,000 mg/kg, respectively). Six treatments were prepared and evaluated during the ripening process and storage time: C1 (control with 150 mg/kg sodium nitrite and 150 mg/kg sodium nitrate), C2 (control without sodium nitrite/nitrate), R05 (0.5% radish powder), R1 (1% radish powder), B05 (0.5% beetroot powder) and B1 (1% beetroot powder). The addition of vegetable powders influenced moisture content, weight loss and water activity of sausages. Nitrite was formed from radish and beetroot powders during the ripening process, especially in R1 and B1 treatments. Beetroot powder affected colour, pigments and lactic acid bacteria counts. The results of pH, colour, lipid oxidation, nitrite and nitrate analysis suggest R1 treatment as a potential nitrite replacer obtained from a simple and feasible drying process.

Production and characterization of promising ß-stable seed crystals to modulate the crystallization of fat-based industrial products.

Spray cooling or spray chilling is a technique for obtaining solid lipid microparticles (SLMs) within the diameter range in micrometers using low temperatures and no organic solvents. It is a low-cost technique and is easy to scale-up. The production of SLMs into ß-form represents a technological challenge due to the fast crystallization given by the spray cooling system, which generally results in SLMs crystallized into the metastable polymorphic form α. This study focuses on the production and characterization of SLMs by spray cooling using hard fat soybean oil (HS) added of D-limonene or canola oil, aiming to their application as ß-seed crystals into lipid systems. The ß-seed crystals could turn into an alternative lipid material to be used in fat-based products that present the preferential ß' crystallization, like palm oil, increasing its compatibility with cocoa butter (CB) and allowing for the development of substitutes. The obtained SLMs showed spherical geometry and no agglomeration during storage at 25 °C for up to 30 days, verified by scanning electron microscopy (SEM). The mean diameters (D50) were between 150 and 200 µm and the ß' and ß-form, determined by X-ray diffraction (XRD), appeared immediately after the crystallization process by spray cooling using HS added of 5% D-limonene (the HS control sample presented only the α-form). The SLMs of this study demonstrated their potential use as ß-seed crystals into lipid systems.

Influence of pulp on the microbial diversity during cupuassu fermentation.

Cupuassu (Theobroma grandiflorum Schum) is a fruit belonging to the same genus as cocoa and, through seed fermentation, a chocolate-like product called "the cupulate" is obtained. The pulp is removed from the seeds before fermentation because its abundance hinders the process. Unlike cocoa, little is known about the microbial diversity involved in cupuassu fermentation. The goal of this study was to explore the use of next-generation sequencing to identify the yeasts and bacteria communities involved in cupuassu seed fermentation on three different pulp concentrations (0, 7.5, and 15%) as well as two turning schemes on the microbial growth. In order to do that, a massive sequencing of the 16S and ITS4 rRNA region (S) using the Illumina MiSeq Platform identified some genera of bacteria and yeasts, respectively, in the fermentation environment. Taxonomic analyses of both communities, especially at the genus level, revealed a predominance of yeasts such as Pichia and Hanseniaspora, and bacteria such as Acetobacter and Lactobacillus. A predominance of bacteria over yeasts diversity was observed in the experiments with higher pulp concentrations (15%). The physicochemical analysis showed that fermentation of samples with 15% pulp exhibited longer fermentation times, the highest temperatures, and elevated production of organic acids such as acetic acid, a precursor of flavor. In addition, the turning applied every 24 h to the mass slightly favored the formation of flavor precursors. It seems that the abundance and composition of cupuassu pulp, rich in organic compounds, can influence the diversity of some populations of yeasts. Some of those compounds identified in previous studies are terpenes with antimicrobial activity. More studies will be necessary to confirm if the presence of terpenes compounds in the cupuassu pulp exert some inhibitory action on microorganism diversity.

Mathematical modeling of uvaia byproduct drying and evaluation of quality parameters.

Uvaia (Eugenia pyriformis) frozen pulp processing generates a solid byproduct that can potentially contain important components of human nutrition. In this study, the drying of uvaia byproduct was studied. Two different drying treatments were tested: drying of wet waste and drying of waste with prior removal of water by centrifugation. Three drying temperatures were used: 40, 60, and 80 °C. Eight models were applied to fit the drying curves: Page, Lewis, Modified Page, Logarithmic, Midilli, Wang and Singh, Henderson and Pabis, and Weibull. Midilli presented an excellent fit to the curves. The effective moisture diffusivity of the uvaia byproduct ranged between 8.52 × 10-10 and 3.22 × 10-9 m2/s. The activation energy was 25.65 and 24.97 kJ/mol for non-centrifuged and centrifuged assays, respectively. The dried byproducts had a reduction of 3-21% of the total phenolic content against the control. The assay performed at 40 °C with centrifugation presented the lowest total color difference value.

Differentiation of cocoa nibs from distinct origins using comprehensive two-dimensional gas chromatography and multivariate analysis.

Headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography was used for identification of volatiles compounds responsible for differentiation between cocoa nibs from Brazil and Ivory Coast. The unfolded GC×GC with Flame Ionization Detector (FID) chromatograms were first aligned using the correlation optimization warping (COW) algorithm and normalized. After that, Fisher ratio was calculated for each variable, and a threshold value was chosen to select the variables that best promote a separation of nibs samples from different sources in a principal component analysis (PCA) model. To identify the relevant compounds for the separation, representative samples of each source were analyzed in the same conditions by GC×GC with detection by quadrupole mass spectrometry. Finally, the average peak volumes for each key compound obtained for both classes were compared using a Student t-test and it was possible to identify 15 volatile compounds responsible for differentiation between cocoa nibs from Brazil and Ivory Coast.

Formation of volatile compounds during cupuassu fermentation: Influence of pulp concentration.

Cupuassu (Theobroma grandiflorum Schum) is a native fruit of the Amazon region and from its seeds fermentation, it is possible to obtain a product similar to chocolate, known as cupulate. The aim of this study was to evaluate the influence of the pulp concentration on the formation of volatile compounds during fermentation. Considering the high quantity of pulp that naturally involves the seeds (38%), which represents an obstacle for fermentation, two different fermentation experiments were conducted: with seeds totally (0%) and partially (15%) depulped. Seeds collected during and after fermentation, after drying (beans), after roasting and deshelling (nibs) and cupulates (obtained using the same methods of chocolate processing) were analyzed through identification and relative quantification by GC-MS in order to check the profile of volatile compounds formed. Results showed that the depulping implies in considerable reduction of important volatile compounds. A wider diversity of volatile compounds such as aldehydes, ketones and alcohols were found in the experiment with 15% pulp during fermentation. Tetramethylpyrazine was the unique pyrazine found in the cupulate samples that had significant difference (p<0.05) among the samples in its concentration. Among the others compounds identified, it is important to emphasize the presence of compounds with fruity and floral notes, generally present in fine/flavor cocoa. This study demonstrated the importance of the pulp in the development of important compounds responsible for the desirable flavor.

Plant and metagenomic DNA extraction of mucilaginous seeds.

The pulp surrounding the seeds of some fruits is rich in mucilage, carbohydrates, etc. Some seeds are rich in proteins and polyphenols. Fruit seeds, like cacao (Theobroma cacao) and cupuassu (Theobroma grandiflorum), are subjected to fermentation to develop flavor. During fermentation, ethanol is produced [2-6]. All of these compounds are considered as interfering substances that hinder the DNA extraction [4-8]. Protocols commonly used in the DNA extraction in samples of plant origin were used, but without success. Thus, a protocol for DNA samples under different conditions that can be used for similar samples was developed and applied with success. The protocol initially described for RNA samples by Zeng et al. [9] and with changes proposed by Provost et al. [5] was adapted for extracting DNA samples from those described. However, several modifications have been proposed:•Samples were initially washed with petroleum ether for fat phase removal.•RNAse was added to the extraction buffer, while spermidin was removed.•Additional steps of extraction with 5 M NaCl, saturated NaCl and CTAB (10%) were included and precipitation was carried out with isopropanol, followed by washing with ethanol.

Behavior of Salmonella during fermentation, drying and storage of cocoa beans.

Due to cocoa being considered a possible source of Salmonella contamination in chocolate, the behavior of Salmonella during some cocoa pre-processing stages (fermentation, drying and storage) was investigated. The fermentation process was carried out on a pilot scale (2 kg beans/box) for 7 days. Every day a fermentation box was inoculated with a Salmonella pool (ca. 4 log MPN/g). The results showed that Salmonella did not affect (P>0.05) the growth of the main microorganism groups involved in cocoa fermentation. On the other hand, the pathogen was influenced (P<0.05) by yeast, acetic acid bacteria and pH. In spite of Salmonella showing counts ≤ 1 log MPN/g in the first days, at the end of fermentation it grew in all samples, reaching counts as high as 7.49 log MPN/g. For drying and storage, cocoa beans were inoculated during the fermentation (experiment A) or during the drying (experiment B). In these stages the decline of the water activity affected the pathogen behavior. In experiment A during the drying, Salmonella count increased in most of the samples. In experiment B either a slight growth or no growth in the samples inoculated up to 48 h was observed, whereas the other samples showed reductions from the initial count. After 30 days of storage at room temperature, the water activity decreased to 0.68, and reductions of Salmonella ranged from 0.93 to 2.52 log MPN/g. Despite the reductions observed during the storage, the pathogen was detected even after 120 days. Therefore, the results showed that Salmonella growth or survival depends on when the contamination occurs.

Occurrence of ochratoxin A in cocoa by-products and determination of its reduction during chocolate manufacture.

This work reports an investigation carried out to assess the natural occurrence of ochratoxin A in 168 samples from different fractions obtained during the technological processing of cocoa (shell, nibs, liquor, butter, cake and cocoa powder) and the reduction of ochratoxin A during chocolate manufacture. Ochratoxin A analyses were performed with immunoaffinity columns and detection by high performance liquid chromatography. Concerning the natural ochratoxin A contamination in cocoa by-products, the highest levels of ochratoxin A were found in the shell, cocoa powder and cocoa cake. The cocoa butter was the least contaminated, showing that ochratoxin A seems to remain in the defatted cocoa solids. Under the technological conditions applied during the manufacture of chocolate in this study and the level of contamination present in the cocoa beans, this experiment demonstrated that 93.6% of ochratoxin A present in the beans was reduced during the chocolate producing.

Inactivation of Salmonella during cocoa roasting and chocolate conching.

The high heat resistance of Salmonella in foods with low water activity raises particular issues for food safety, especially chocolate, where outbreak investigations indicate that few colony-forming units are necessary to cause salmonellosis. This study evaluated the efficiency of cocoa roasting and milk chocolate conching in the inactivation of Salmonella 5-strain suspension. Thermal resistance of Salmonella was greater in nibs compared to cocoa beans upon exposure at 110 to 130°C. The D-values in nibs were 1.8, 2.2 and 1.5-fold higher than those calculated for cocoa beans at 110, 120 and 130°C. There was no significant difference (p>0.05) between the matrices only at 140°C. Since in the conching of milk chocolate the inactivation curves showed rapid death in the first 180 min followed by a lower inactivation rate, and two D-values were calculated. For the first time interval (0-180 min) the D-values were 216.87, 102.27 and 50.99 min at 50, 60 and 70°C, respectively. The other D-values were determined from the second time interval (180-1440 min), 1076.76 min at 50°C, 481.94 min at 60°C and 702.23 min at 70°C. The results demonstrated that the type of matrix, the process temperature and the initial count influenced the Salmonella resistance.

Fermentation of cacao (Theobroma cacao L.) seeds with a hybrid Kluyveromyces marxianus strain improved product quality attributes.

Fermentation of Theobroma cacao (cacao) seeds is an absolute requirement for the full development of chocolate flavor precursors. An adequate aeration of the fermenting cacao seed mass is a fundamental prerequisite for a satisfactory fermentation. Here, we evaluated whether a controlled inoculation of cacao seed fermentation using a Kluyveromyces marxianus hybrid yeast strain, with an increased pectinolytic activity, would improve an earlier liquid drainage ('sweatings') from the fermentation mass, developing a superior final product quality. Inoculation with K. marxianus increased by one third the volume of drained liquid and affected the microorganism population structure during fermentation, which was detectable up to the end of the process. Introduction of the hybrid yeast affected the profile of total seed protein degradation evaluated by polyacrylamide gel electrophoresis, with improved seed protein degradation, and reduction of titrable acidity. Sensorial evaluation of the chocolate obtained from beans fermented with the K. marxianus inoculation was more accepted by analysts in comparison with the one from cocoa obtained through natural fermentation. The increase in mass aeration during the first 24 h seemed to be fundamental for the improvement of fermentation quality, demonstrating the potential application of this improved hybrid yeast strain with superior exogenous pectinolytic activity.

1H NMR, a rapid method to monitor organic acids during cupuassu (Theobroma grandiflorum Spreng) processing.

The development of an analytical method using 1H nuclear magnetic resonance (1H NMR) spectrometry to monitor cupuassu (Theobroma grandiflorum Spreng) bean fermentation, drying, and roasting processes is reported. The analysis of organic acids and alcohols of crude water extracts of cupuassu ground kernels were monitored by HPLC and 1H NMR spectroscopy. The residual protein signals caused deleterious effects on acid and alcohol quantifications. Therefore, the analytical procedures were optimized by sample cleanup and water suppression pulse sequences in order to obtain compatible data using HPLC and 1H NMR. The quantification of lactic acid, acetic acid, and 2,3-butanediol by NMR is 5- to 10-fold faster than by HPLC, with the advantage of providing the identification of several chemical species in a single experiment. Application of these analytical conditions to some cupuassu samples revealed that this methodology can be applied to the quality profiles of fermentation and roasting processes.